ET1702-52

Actin Recombinant Rabbit Monoclonal Antibody [JF47-01]

cat.: ET1702-52

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat, Zebrafish
Applications:	WB, IHC-P, IF-Cell, FC
Clonality:	Monoclonal
Clone number:	JF47-01

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 42 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human Actin aa 303-349 / 377.
Positive control:	HeLa cell lysate, C2C12 cell lysate, L6 cell lysate, PC-12 cell lysate, COS-1 cell lysate, Zebrafish tissue lysate, Hybrid fish (crucian-carp) brain tissue lysate, Hybrid fish (crucian-carp) kidney tissue lysate, human skeletal muscle tissue, mouse skeletal muscle tissue, rat skeletal muscle tissue, HeLa, NIH/3T3, L6, PC-12.
Subcellular location:	Cytoplasm.
Recommended Dilutions: WB IHC-P IF-Cell FC	1:5,000-1:20,000 1:200 1:250-1:500 1:100-1:1,000
Uniprot #:	SwissProt: P68133 Human \| P68134 Mouse \| P68136 Rat
Alternative names:	a actin ACTA ACTA1 Actin alpha skeletal muscle Actin actin, alpha 1, skeletal muscle 1 actin, alpha 1, skeletal muscle Actin, alpha skeletal muscle actina actine ACTS_HUMAN aktin Alpha Actin 1 Alpha skeletal muscle Actin alpha skeletal muscle alpha-actin Alpha-actin-1 ASMA CFTD CFTD1 CFTDM MPFD NEM1 NEM2 NEM3 nemaline myopathy type 3

Images

	Fig1: Western blot analysis of Actin on different lysates with Rabbit anti-Actin antibody (ET1702-52) at 1/5,000 dilution. Lane 1: HeLa cell lysate Lane 2: C2C12 cell lysate Lane 3: L6 cell lysate Lane 4: PC-12 cell lysate Lane 5: COS-1 cell lysate Lane 6: Zebrafish tissue lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 42 kDa Observed band size: 42 kDa Exposure time: 4 seconds; ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1702-52) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
	Fig2: Western blot analysis of Actin on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1702-52, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature. Positive control: Lane 1: Hybrid fish (crucian-carp) brain tissue lysate Lane 2: Hybrid fish (crucian-carp) kidney tissue lysate
	Fig3: Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue with Rabbit anti-Actin antibody (ET1702-52) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1702-52) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

	Fig4: Immunohistochemical analysis of paraffin-embedded mouse skeletal muscle tissue with Rabbit anti-Actin antibody (ET1702-52) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1702-52) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig5: Immunohistochemical analysis of paraffin-embedded rat skeletal muscle tissue with Rabbit anti-Actin antibody (ET1702-52) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1702-52) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig6: Immunocytochemistry analysis of HeLa cells labeling Actin with Rabbit anti-Actin antibody (ET1702-52) at 1/250 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Actin antibody (ET1702-52) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

	Fig7: Flow cytometric analysis of HeLa cells labeling Actin. Cells were fixed and permeabilized. Then stained with the primary antibody (ET1702-52, 1/100) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
	Fig8: Immunocytochemistry analysis of NIH/3T3 cells labeling Actin with Rabbit anti-Actin antibody (ET1702-52) at 1/250 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Actin antibody (ET1702-52) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
	Fig9: Flow cytometric analysis of NIH/3T3 cells labeling Actin. Cells were fixed and permeabilized. Then stained with the primary antibody (ET1702-52, 1/100) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Fig10: Immunocytochemistry analysis of L6 cells labeling Actin with Rabbit anti-Actin antibody (ET1702-52) at 1/500 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Actin antibody (ET1702-52) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Fig11: Flow cytometric analysis of PC-12 cells labeling Actin.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1702-52, 1/100) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.